Cell Biology


    Protocols in Current Issue
    Isolation and Quantification of Plant Extracellular Vesicles
    Authors:  Brian D. Rutter, Katie L. Rutter and Roger W. Innes, date: 09/05/2017, view: 279, Q&A: 0
    [Abstract] Extracellular vesicles (EVs) play an important role in intercellular communication by transporting proteins and RNA. While plant cells secrete EVs, they have only recently been isolated and questions regarding their biogenesis, release, uptake and function remain unanswered. Here, we present a detailed protocol for isolating EVs from the ...
    Isolation of Cytosol, Microsome, Free Polysomes (FPs) and Membrane-bound Polysomes (MBPs) from Arabidopsis Seedlings
    Authors:  Yonghui Zhao and Shengben Li, date: 08/05/2017, view: 461, Q&A: 0
    [Abstract] The plant endomembrane system plays vital roles for synthesis, modification and secretion of proteins and lipids. From the classic view, only mRNAs encoding secreted proteins could be targeted to the endoplasmic reticulum (ER) for translation via a co-translational translocation manner, however, recently this model has been challenged by ...
    Photometric Assays for Chloroplast Movement Responses to Blue Light
    [Abstract] Assessment of chloroplast movements by measuring changes in leaf transmittance is discussed, with special reference to the conditions necessary for reliable estimation of blue light–activated chloroplast responses.
    Plasma Membrane Preparation from Lilium davidii and Oryza sativa Mature and Germinated Pollen
    Authors:  Ning Yang, Bing Han, Lingtong Liu, Hao Yang and Tai Wang, date: 05/20/2017, view: 798, Q&A: 0
    [Abstract] Pollen germination is an excellent process to study cell polarity establishment. During this process, the tip-growing pollen tube will start elongating. The plasma membrane as the selectively permeable barrier that separate the inner and outer cell environment plays crucial roles in this process. This protocol described an efficient aqueous ...
    Nuclei Isolation from Nematode Ascaris
    Authors:  Yuanyuan Kang, Jianbin Wang and Richard E. Davis, date: 05/05/2017, view: 875, Q&A: 0
    [Abstract] Preparing nuclei is necessary in a variety of experimental paradigms to study nuclear processes. In this protocol, we describe a method for rapid preparation of large number of relatively pure nuclei from Ascaris embryos or tissues that are ready to be used for further experiments such as chromatin isolation and ChIP-seq, nuclear RNA ...
    Physical Removal of the Midbody Remnant from Polarised Epithelial Cells Using Take-Up by Suction Pressure (TUSP)
    Authors:  Miguel Bernabé-Rubio, David C. Gershlick and Miguel A. Alonso, date: 04/20/2017, view: 987, Q&A: 0
    [Abstract] In polarised epithelial cells the midbody forms at the apical cell surface during cytokinesis. Once severed, the midbody is inherited by one of the daughter cells remaining tethered to the apical plasma membrane where it participates in non-cytokinetic processes, such as primary ciliogenesis. Here, we describe a novel method to physically remove ...
    Isolation of Exosomes from Semen for in vitro Uptake and HIV-1 Infection Assays
    Authors:  Marisa N. Madison, Jennifer L. Welch and Chioma M. Okeoma, date: 04/05/2017, view: 1034, Q&A: 0
    [Abstract] Exosomes are membranous extracellular nanovesicles of endocytic origin. Exosomes are known to carry host and pathogen-derived genomic, proteomic, lipidomic cargos and other extraneous molecules. Exosomes are secreted by diverse cell types into the extracellular milieu and are subsequently internalized by recipient neighboring or distal cells. Upon ...
    Differential Salt Fractionation of Nuclei to Analyze Chromatin-associated Proteins from Cultured Mammalian Cells
    Authors:  Christin Herrmann, Daphne C. Avgousti and Matthew D. Weitzman, date: 03/20/2017, view: 1506, Q&A: 0
    [Abstract] Nucleosomes are the core units of cellular chromatin and are comprised of 147 base pairs (bp) of DNA wrapped around an octamer of histone proteins. Proteins such as chromatin remodelers, transcription factors, and DNA repair proteins interact dynamically with chromatin to regulate access to DNA, control gene transcription, and maintain genome ...
    Ca2+ Measurements in Mammalian Cells with Aequorin-based Probes
    Authors:  Anna Tosatto, Rosario Rizzuto and Cristina Mammucari, date: 03/05/2017, view: 1025, Q&A: 0
    [Abstract] Aequorin is a Ca2+ sensitive photoprotein suitable to measure intracellular Ca2+ transients in mammalian cells. Thanks to recombinant cDNAs expression, aequorin can be specifically targeted to various subcellular compartments, thus allowing an accurate measurement of Ca2+ uptake and release of different ...
    Quantitative Analysis of Exosome Secretion Rates of Single Cells
    Authors:  Yu-Jui Chiu, Wei Cai, Tiffany Lee, Julia Kraimer and Yu-Hwa Lo, date: 02/20/2017, view: 1682, Q&A: 0
    [Abstract] To study the inhomogeneity within a cell population including exosomes properties such as exosome secretion rate of cells and surface markers carried by exosomes, we need to quantify and characterize those exosomes secreted by each individual cell. Here we develop a method to collect and analyze exosomes secreted by an array of single cells using ...